AChE/BChE-IN-23 (Compound 6e) is an AChE/BChE inhibitor (IC50 for AChE, eqBChE and hBChE are 0.91 μM, 1.19 μM and 1.01 μM, respectively). AChE/BChE-IN-23 has antioxidant effects and inhibits Aβ1-42 and Tau protein aggregation. AChE/BChE-IN-23 also inhibits microglial activation by inhibiting reactive oxygen species release and mitochondrial damage. AChE/BChE-IN-23 also reduces NLRP3 inflammasomes in human microglia. It also reverses scopolamine-induced memory impairment in mice.

Physicochemical Properties

Molecular Formula	C19H21N5O3
Molecular Weight	367.40
Appearance	Typically exists as solids at room temperature
HS Tariff Code	2934.99.9001
Storage	Powder-20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month
Shipping Condition	Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

Biological Activity

Targets	hAChE 0.91 μM (IC50); eqBCHE 1.19 μM (IC50); NLRP3 hBCHE 1.02 μM (IC50)
ln Vitro	AChE/BChE-IN-23 causes propidium iodide (with affinity for AChE enzyme) to shift (5 μM, 2.06%; 10 μM, 11.78%; 20 μM, 14.47%; 50 μM, 27.53%) [1]. AChE/BChE-IN-23 has antioxidant effect (20 μM; DPPH scavenging rate is 47.30%, IC50:15.17 μM) [1]. AChE/BChE-IN-23 (3.125 μM, 72 h) can effectively inhibit the aggregation of Aβ1-42 (12.5 μM) [1]. AChE/BChE-IN-23 (1.125 μM, 72 h) can effectively inhibit the aggregation of Tau (5 μM) [1]. AChE/BChE-IN-23 (1-20 μM, 24 h) has no toxicity to PC-12 cells[1]. AChE/BChE-IN-23 (12.5 μM, 24 h) inhibits microglial activation by reducing the release of ROS and restoring mitochondrial membrane potential[1]. AChE/BChE-IN-23 (12.5 μM, 24 h) exerts anti-inflammatory effects by inhibiting the activation of inflammasomes and NF-κB in microglia[1].
ln Vivo	AChE/BChE-IN-23 (10 mg/kg, intraperitoneal injection, 14 days) reversed scopolamine-induced Alzheimer's disease by enhancing spatial and cognitive memory in AD mice[1]. AChE/BChE-IN-23 (2000 mg/kg, gavage, 14 days) showed no toxicity to Swiss albino mice, including weight gain, behavioral abnormalities, convulsions, tremors, or diarrhea associated with acute toxicity [1].
Cell Assay	Western Blot Analysis[1] Cell Types: HMC-3 cells Tested Tested Concentrations: 12.5 μM Incubation Duration: 24 h Experimental Results: Suppressed NLRP3, caspase-1, IL-1β and IL-18 expression. Immunofluorescence[1] Cell Types: HMC-3 cells Tested Tested Concentrations: 12.5 μM Incubation Duration: 24 h Experimental Results: Reduced mitochondrial ROS and stabilizes the mitochondrial membrane potential.
Animal Protocol	Animal/Disease Models: Scopolamine (1 mg/kg) induced Alzheimer’s disease (AD) model; 6-week-old Swiss albino mice[1] Doses: 10 mg/kg Route of Administration: Intraperitoneal injection (i.p.), 14 days Experimental Results: Decreased escape latency, distance traveled in Morris Water Maze (MWM) experiment. Increased latency period in passive avoidance. Increased BDNF and Nr4a2 in brain.
References	[1]. Discovery of novel substituted (Z)-N'-hydroxy-3-(3-phenylureido)benzimidamide derivatives as multifunctional molecules targeting pathological hallmarks of Alzheimer's disease. Eur J Med Chem. 2024;280:116959.

Solubility Data

Solubility (In Vitro)

May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples

Solubility (In Vivo)

Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples. Injection Formulations (e.g. IP/IV/IM/SC) Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] *Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline) Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline) Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline) Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil) Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Oral Formulations Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). Oral Formulation 3: Dissolved in PEG400 Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose Oral Formulation 6: Mixing with food powders Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	2.7218 mL	13.6091 mL	27.2183 mL
	5 mM	0.5444 mL	2.7218 mL	5.4437 mL
	10 mM	0.2722 mL	1.3609 mL	2.7218 mL

*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.